Construction of phage antibody library using sensitized in vitro B-lymphocytes of liver cancer patients
- VernacularTitle:用体外致敏联合EB病毒转化技术构建抗肝癌的人源Fab噬菌体抗体库
- Author:
Xuan SHUI
;
Guancheng LI
;
Yuehui LI
;
Jian HUANG
;
Yan ZHAO
- Publication Type:Journal Article
- Keywords:
EBV immortalization;
phage antibody library;
liver cancer;
Fab fragment
- From:
Journal of Xi'an Jiaotong University(Medical Sciences)
1982;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct human phage antibody library against hepatoma carcinoma.Methods Peripheral blood mononuclear cells(PBMCs) of patients with liver cancer were sensitized in vitro and transformed by Epstein-Barr virus(EBV).The genes of light chain and Fd of antibodies were amplified by RT-PCR.Fab genes were cloned into vector pComb3 and transformed into E.coli XLI-Blue by electroporation to construct the Fab-displaying phage antibody library.Results ELISA detection showed that 4 liver cancer patients' B cells transformed by EBV could produce specific antibodies to hepatoma carcinoma cell.Totally 13 types of light chain genes and 28 types of Fd genes were obtained by RT-PCR.The capacity of the primary phage library was 1.7?107pfu/mL.The percentage of recombinant clones was about 100%.Conclusion A human phage antibody library has been constructed successfully by means of EBV transformation technique.
